Ultrasonographic examination of the forestomachs and the abomasum in ruminal drinker calves

Background

The study investigated the ultrasonographic appearance of the reticulum, rumen, omasum and abomasum of calves with ruminal drinking syndrome.

Methods

In ten milk-fed calves with ruminal drinking syndrome the reticulum, rumen, omasum and abomasum were examined by ultrasonography using a 5-MHz linear transducer before, during and after the ingestion of milk.

Results

The reticulum could be imaged in eight of ten calves before feeding. The reticular wall appeared as an echoic line, similar to mature cattle, and reticular folds were seen in eight calves. The reticular content appeared as echoic heterogeneous fluid. Reticular contractions were biphasic with 1.0 ± 0.38 contractions per minute. The rumen had a mean wall thickness of 2.1 mm dorsally, 3.5 mm at the level of the longitudinal groove, and 3.2 mm ventrally. The ventral sac of the rumen of all calves contained echoic heterogeneous liquid. During feeding the milk entering the rumen could be seen as hyperechoic liquid in five calves. The omasum was seen on the right side as a crescent-shaped line medial to the liver in seven calves. Only the omasal wall closest to the transducer was seen as an echoic line with a mean thickness of 2.7 mm. The ultrasonographic appearance of the omasum did not change during or after feeding. The abomasum was seen immediately caudal to the xyphoid on both sides of the midline before feeding. The mean length at the ventral midline was 22.2 cm. The ingesta were heterogeneous in all calves and the abomasal folds were distinct in eight. The mean lateral expansion of the abomasum from the ventral midline to the left and right varied from 8.7 to 13.8 cm and from 4.3 to 11.3 cm. The milk entering the abomasum was observed in all calves, and signs of milk clotting were seen in all calves 15 minutes after feeding.

Conclusion

This study showed that ultrasonography is useful for detecting milk in the reticulum and rumen of calves with ruminal drinking syndrome.     

Ultrasonography of the reticulum, rumen, omasum and abomasum before, during and after ingestion of hay and grass silage in 10 calves

The reticulum, rumen, omasum and abomasum were assessed via ultrasonography in 10 healthy female calves before, during and 2h after feeding hay and grass silage. The evaluations were made using an ultrasound machine with a 5.0MHz linear transducer. The reticulum could be visualized before feeding in all the calves. Its appearance and pattern of contractions were similar to those in adult cattle, although the amplitude (5.2±1.06cm) and velocity (3.5±1.42cm/s) of the first contraction were markedly less than in adult cattle. The position and size of the entire rumen including the dorsal and ventral sacs and the ruminal contents were assessed. Except for its smaller size, the ultrasonographic appearance of the omasum of calves was similar to that of adult cattle. The abomasum was seen to the left and right of the ventral midline before feeding in all calves; it occupied considerably more space on the left than the right. Compared with its appearance before feeding, the ultrasonographic appearance of the rumen, omasum and abomasum did not change during or after feeding. Ultrasonography is an ideal imaging tool for evaluating the reticulum, rumen, omasum and abomasum before, during and after feeding in calves.     

Ultrasonography of the reticulum, rumen, omasum, and abomasum in 10 calves before, during, and after ingestion of milk

The reticulum, rumen, omasum, and abomasum were assessed via ultrasonography before, during, and 15, 30, and 120 minutes after feeding milk to 10 healthy calves. The ultrasonographic examinations were conducted using a 5.0 MHz linear transducer. Loops were recorded on video for further evaluation. The reticulum could be visualised before feeding in seven calves. Its appearance and pattern of contractions were similar to those in adult cattle, although the amplitude (1.7 ± 0.75 cm) and velocity (2.7 ± 1.34 cm/s) of the first contraction were smaller than in adult cattle. The reticulum could not be visualised in any of the calves during feeding as it was displaced cranially and laterally and therefore being obscured by the lungs as the abomasum expanded with the ingested milk. 2 hours post ingestion it remained obscured in 5 individual and was visualized again the other 5. The position and size of the entire rumen including the dorsal and ventral sacs and the ruminal contents were assessed. There were no changes in the ultrasonographic appearance of the rumen during or after feeding. Except for its smaller size, the ultrasonographic appearance of the omasum of calves was similar to that of adult cattle. Milk flow through the omasum could not be seen in any of the calves, and there were no changes in the appearance of the omasum during and after feeding. The abomasum was seen to the left and right of the ventral midline before feeding in all calves; it occupied considerably more space on the left than the right. The flow of milk into the abomasum and milk clotting, which occurred 15 minutes after feeding, could be seen in all calves. The milk clots started to slowly disintegrate 30 minutes after the start of feeding, and by 2 hours post feeding, this process was greatly advanced but remained incomplete. Ultrasonography is an ideal tool for the evaluation of the reticulum, rumen, omasum, and abomasum before, during, and after the ingestion of milk in calves.     

Provision of beta‐glucan prebiotics (cellooligosaccharides and kraft pulp) to calves from pre‐ to post‐weaning period on pasture

We conducted two feeding experiments to evaluate the effects of supplementation with either cellooligosaccharide or kraft pulp on growth performance in grazing beef calves (Japanese Black) from 4 weeks pre‐weaning to 12 to 16 weeks post‐weaning. In Experiment 1 (20‐week duration), nine calves (2.9‐month‐old females) were assigned to either a control group (CON) or an experimental group (CEL) fed cellooligosaccharide at a rate of 10 g/day mixed with concentrate. Average daily weight gain tended to be greater in CEL than in CON, especially after 1 month of weaning. In Experiment 2 (16‐week duration), 10 calves (2.0‐month‐old females) were assigned to either a control group or an experimental group (KRA) fed kraft pulp at a rate of 10% replacement of total digestible nutrients with concentrate. The proportion of fibrolytic bacteria increased and that of methanogenic Archaea decreased in the rumen microbial community composition of KRA calves in Experiment 2, whereas the decrease in Fibrobacter and Archaea was observed in CEL calves at first 4 weeks in Experiment 1. We conclude that beta‐glucan prebiotic supplementation to grazing calves at pre‐weaning would affect rumen microbial composition and modified rumen fermentation characteristics, leading to a better rumen environment via different means.     

Expression of Proton-dependent Oligopeptide Transporters (POTs) in Calves' Tissues

This test was designed to study the expression of POTs (PepT1, PepT2, PHT1 and PHT2) mRNA in calves'tissues.Tissue-specific expression of the mRNA corresponding to peptide transporter protein in four 3-month-old Chinese Holstein calves was examined by relative quantitative RT-PCR analysis.The results showed that the expression of PepT1 mRNA in the rumen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PepT2 mRNA in the liver and kidney was extremely significantly higher than that in the heart, spleen, thymus, muscle, rumen, reticulum, omasum and abomasum (P<0.01);The expression of PHT1 mRNA in the lung and spleen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PHT2 mRNA in the lung and thymus was significantly higher than that in the heart, liver, kidney, spleen, muscle, rumen, reticulum, omasum and abomasum (P<0.05).The results indicated that PepT1, PepT2, PHT1 and PHT2 mRNA were respectively abundantly expressed in rumen, kidney and liver, spleen and lung, lung and thymus.     

犊牛组织中小肽转运体(POTs)的表达研究

为研究犊牛部分组织中小肽转运体(proton-dependent oligopeptide transporters,POTs) (PepT1、PepT2、PHT1、PHT2) mRNA的表达,试验选用4头3月龄的中国荷斯坦犊牛进行屠宰,采用实时荧光定量PCR方法来定量组织中小肽转运蛋白表达水平。结果表明,犊牛PepT1 mRNA在瘤胃中表达丰度极显著高于心脏和肌肉(P<0.01);犊牛PepT2 mRNA在肝脏和肾脏中表达丰度极显著高于心脏、脾脏、胸腺、肌肉、瘤胃、网胃、瓣胃、皱胃(P<0.01);犊牛PHT1 mRNA在肺脏和脾脏中表达丰度极显著高于心脏和肌肉(P<0.01);犊牛PHT2 mRNA在肺脏和胸腺中表达丰度显著高于心脏、肝脏、肾脏、脾脏、肌肉、瘤胃、网胃、瓣胃、皱胃(P<0.05)。综上所述,PepT1、PepT2、PHT1、PHT2 mRNA分别在犊牛瘤胃、肾脏和肝脏、脾脏和肺脏、肺脏和胸腺中表达量最高。     

Growth of rumen papillae in weaned calves is associated with lower expression of insulin‐like growth factor‐binding proteins 2, 3, and 6

This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin‐like growth factor‐binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real‐time PCR, and the presence of insulin‐like growth factors I and II (IGF‐I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk‐continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk‐continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk‐continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF‐I and IGF‐II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.     

Severe hypoplasia of the omasal laminae in a Japanese Black steer with chronic bloat--a case report

An 11-month-old Japanese Black steer with chronic bloat underwent clinical and histological analyses. During the observation period, it showed normal appetite and fecal volume but persistent chronic bloat symptoms. Compared to controls, the steer's feces contained undigested large straws. Necropsy revealed normal rumen, reticulum, and abomasum but a small omasum. The rumen, reticulum, and abomasum mucosa was normal, with well-developed ruminal papillae. However, severe hypoplasia of the omasal laminae was observed along with hypoplasia reticular groove and ruminoreticular fold. The contents of the reticulum, omasum, and abomasums comprised undigested large sized hay particles. The omasum papillae showed no pathological abnormalities. This is a rare case of a steer with chronic bloat probably caused by severe hypoplasia of the omasal laminae.     

A comparative histological study of the number and size of the myenteric ganglia and neurones in the fore-stomach and abomasum of grey, white and black Karakul lambs.

Homozygous grey Karakul lambs are born with a lethal genetic factor responsible for death at weaning age. When put on a high roughage diet under field conditions they develop distended, thin-walled rumens and sand impacted abomasa. Homozygous white Karakul lambs have a similar factor but survive for a longer period. Black Karakul lambs are not affected. The present study was undertaken to compare by image analysis the number and size of the myenteric ganglia, and the number of myenteric neurones in the walls of the fore-stomach and abomasum of 24-h-old grey, white and black Karakul lambs. One square centimetre samples were taken from analogous areas of the rumen, reticulum, omasum and abomasum of 38 embalmed Karakul lambs. Haematoxylin and eosin stained histological sections of each sample were studied with a Vids 2 Image Analyzer. One way analysis of variance indicated a significant difference between the groups regarding the number and size of the myenteric ganglia and in the number of myenteric neurones in the reticulum, rumen and abomasum. The number and size of the ganglia and the number of neurones was greatest in the black lambs and decreased progressively in the white and grey lambs. The omasum was not affected. It is suggested that the paucity of myenteric ganglia and neurones in the regions examined is instrumental in causing the lethal condition described above.     

Morphological study on the stomach of the lesser mouse deer (Tragulus javanicus) with special reference to the internal surface.

The stomach of the lesser mouse deer (Tragulus javanicus) was observed macroscopically. It consisted of only three compartments, rumen, reticulum and abomasum without omasum. The rumen was S-shaped with large ventral and caudoventral blind sacs and the reticulum was larger than the abomasum. Internally, the rumen was covered with numerous ruminal papillae even on the pillars and the ruminoreticular fold. These papillae were leaf- or tongue-like shaped and varied in size and density. The reticulum had honey-combed crests and the secondary crests were found rarely. The lips of the reticular groove were prominent and more developed in the aboral part than in the oral one. A sac-like transition zone, which had more prominent mucosal folds than had the floor of the reticular groove, was observed between the caudal end of the reticular groove and the abomasum. Mucosal folds of the abomasum were spiral, low but rather thick. These findings were discussed in view of comparison with other ruminants and of possible functional implications.     

Physical characteristics of gastrointestinal content of llama (Lama glama)

Changes in digesta dry matter (DM) and mean digesta particle size (MPS) along the gastrointestinal tract are well known in ruminants, but not in camelids. We collected digesta from the dorsal (d) and ventral (v) first forestomach compartment (C1), the second forestomach compartment (C2), three proximal segments and the subsequent glandular part of the third compartment (C3A‐D), the caecum and the faeces twelve llamas (Lama glama). DM analysis indicates the presence of digesta stratification in the C1, the presence of fluid in the C2 to facilitate the sorting function of this compartment, the fluid‐absorbing function of the proximal parts of the C3, the secretion of enzymes and digestive acids in the C3D, and the water‐resorbing function of the lower intestinal tract. These findings illustrate the functional resemblance between the gastrointestinal tract of camelids and cattle‐like ruminants (C1 equivalent to the rumen with stratified contents, C2 to the reticulum, C3A/B/C to the omasum and C3D to the abomasum). MPS analysis revealed a progressive reduction in MPS from the C1 to the distal C3. This gradual transition is different from the clear‐cut threshold in ruminants between the reticulum and the omasum and had so far only been described in dromedaries (Camelus dromedarius). These findings indicate that regardless of the convergent property of rumination and resemblance of general mechanisms involved in contents stratification and particle sorting, differences between ruminants and camelids exist that could be interpreted as a more efficient functionality of the ruminant forestomach.     

哺乳期藏羊羔羊复胃的发育性变化

为了研究羔羊复胃的发育性变化,试验以0-112日龄哺乳期藏羊羔羊为研究对象,采用大体解剖学方法测定了放牧条件下0、3、7、14、21、28、42、56、72、84、98和112日龄羔羊瘤胃、网胃、瓣胃、皱胃、前胃、总胃重量及各胃室容积的变化情况。结果表明：藏羊羔羊在0-112日龄的各胃室发育迅速,各胃室增重明显,容积显著增加,尤其瘤胃的增重明显;在藏羊生产实践中,70日龄断奶较适宜,56日龄是可接受的最早断奶日龄。     

Effects of Saccharomyces cerevisiae supplementation on growth performance,plasma metabolites and hormones,and rumen fermentation in Holstein calves during pre‐ and post‐weaning periods

This study aimed to evaluate the effects of supplementing Saccharomyces cerevisiae (SC) during the pre‐ and post‐weaning periods on growth, metabolic and hormonal responses, and rumen fermentation in calves. Three‐week‐old Holstein calves were assigned to either control (n = 12) or SC group (n = 12), the latter of which received 2 × 10⁹ cfu/day of SC. The experiment was conducted over a period of 7 weeks around weaning. Daily gain (DG) in the SC group was higher (p < .05) than that in the control group. In the SC group, plasma glucose, insulin, and growth hormone (GH) concentrations were higher (p < .05) and concentrations of glucagon and insulin‐like growth factor 1 (IGF‐1) tended to be higher (p < .1) than in the control group. Proportion of rumen propionate and concentration of rumen ammonia nitrogen at 10 weeks of age were greater (p < .05) in the SC group than that in the control group. Supplementation of SC around weaning may improve dietary nutrient and energy availability and increase plasma GH and IGF‐1 concentrations. These changes observed in SC‐supplemented calves could be closely related to the improvement of DG.     

Uptake of milk with and without solid feed during the monogastric phase: Effect on fibrolytic and methanogenic microorganisms in the gastrointestinal tract of calves

Microbial communities are affected by diet and play a role in the successful transition from milk to a solid diet. The response of microorganisms in the gastrointestinal tract of Holstein bull calves to the uptake of milk with solid feed (control treatment; CT), or milk without solid feed (milk‐only treatment; MT) during the first 3 weeks of life was investigated. Samples were collected from the rumen (fluid and tissue), abomasum (fluid), cecum (fluid and tissue) and feces at 7, 14 and 20 days of age. Calf weight was higher on days 14 and 20 in the MT than the CT. In the rumen at 14 days, the fibrolytic bacteria Fibrobacter succinogenes and Prevotella ruminicola increased in the CT and Ruminococcus flavefaciens increased in the MT. This suggests that R. flavefaciens is not strictly fibrolytic and that it might use milk as a substrate or other microbial species might supply a substrate. Diet affected methanogens, but this may have been due to an indirect effect via an association with Geobacter spp. or other syntrophic partners. The treatments also affected microorganisms in the abomasum, cecum and feces. Our results contribute to an understanding of diet, microbes in the gastrointestinal tract and weaning.     

Effects of forage offering method on performance,rumen fermentation,nutrient digestibility and nutritional behaviour in Holstein dairy calves

The potential effect of dietary forage supplementation on the performance and rumen development in dairy calves is well established. However, limited research has been directed to the comparative effects of forage offering methods on calf performance. The objective of the present study was to determine the effects of forage provision methods (total mixed ration or free choice) on the performance, nutrient digestibility, rumen fermentation and nutritional behaviour in newborn calves. Forty‐five Holstein dairy calves (3 days of age and 41 ± 2 kg of body weight) were assigned to the following three groups (n = 15): (i) starter without forage provision (CON), (ii) starter supplemented with 10% alfalfa hay (AH) as a total mixed ration (AH‐TMR) and (iii) starter and AH as a free‐choice provision (AH‐FC) for a period of 70 days. All the calves were offered 5 l of milk/day from day 3 to 50, and 2.5 l/day from day 50 until weaning on day 56. Dry matter intake (DMI) was greater (p < 0.01) in the AH‐TMR and AH‐FC treatments than in the CON during the pre‐ and post‐weaning periods. Calves fed the AH‐FC diet showed the highest post‐weaning DMI among the treatments. The calves receiving ad libitum forage tended (p = 0.08) to increase crude protein digestibility and overall volatile fatty acids (VFA) concentrations in the rumen. No differences were observed among the treatments at the time spent on standing, lying, eating and performing non‐nutritive oral behaviours. Compared to CON calves, animals in the AH‐TMR treatment spent more time (p < 0.05) ruminating. In conclusion, our data suggest that forage supplementation in both forage offering methods increased total DMI, ruminal pH and ruminating time in dairy calves. Hence, there is no benefit in the free‐choice provision of AH in dairy calves.     

藏绵羊溶菌酶1基因的克隆、原核表达及生物信息学分析

试验旨在研究藏绵羊溶菌酶(lysozyme,LZM)1基因在瘤胃、瓣胃、皱胃、气管、肝脏和肾脏各组织中的表达情况、进化关系及抑菌活性.采用RT-PCR技术克隆藏绵羊LZM1基因,定量PCR检测LZM1基因在瘤胃、瓣胃、皱胃、气管、肝脏和肾脏中的表达情况,并将该基因的氨基酸序列与普通牛胃LZM等氨基酸序列进行比对,根据邻位连接法构建系统进化树;将该基因插入pET-32a质粒后构建得到pET-32a-LZM1重组质粒,转化大肠杆菌BL21(DE3)感受态细胞并诱导表达,SDS-PAGE法鉴定表达产物;采用比浊法测定LZM1蛋白的活性;采用琼脂糖平板扩散法研究异源表达蛋白的抑菌活性.结果表明,从藏绵羊瘤胃、瓣胃、皱胃、气管、肝脏和肾脏各组织中成功克隆藏绵羊LZM1基因,其在瘤胃、瓣胃、皱胃、气管、肝脏和肾脏中均有表达,在皱胃中表达量最高;其氨基酸序列与普通牛胃LZM(GenBank登录号:NM_001080339.1)序列的同源性最高(96%);重组蛋白LZM1分子质量约为35 ku;其比活力约为400 U/mL;该重组LZM1对金黄色葡萄球具有良好的抑菌活性.